A Potent d -Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

Uppalapati, Maruti; Lee, Dong Jun; Mandal, Kalyaneswar; Li, Hongyan; Miranda, Les P.; Lowitz, Joshua; Kenney, John; Adams, Jarrett J.; Ault-Riché, Dana; Kent, Stephen B. H.; Sidhu, Sachdev S.

doi:10.1021/acschembio.5b01006

Cited by 113Chemical Synthesis and AnalysisMonoclonal and Polyclonal Antibodies ResearchAngiogenesis and VEGF in Cancer

A Potent d -Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

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Uppalapati, Maruti, Lee, Dong Jun, Mandal, Kalyaneswar, Li, Hongyan, Miranda, Les P., Lowitz, Joshua, Kenney, John, Adams, Jarrett J. +3 more·2016·ACS Chemical Biology

DOI 10.1021/acschembio.5b01006

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Abstract

Polypeptides composed entirely of d-amino acids and the achiral amino acid glycine (d-proteins) inherently have in vivo properties that are proposed to be near-optimal for a large molecule therapeutic agent. Specifically, d-proteins are resistant to degradation by proteases and are anticipated to be nonimmunogenic. Furthermore, d-proteins are manufactured chemically and can be engineered to have other desirable properties, such as improved stability, affinity, and pharmacokinetics. Thus, a well-designed d-protein therapeutic would likely have significant advantages over l-protein drugs. Toward the goal of developing d-protein therapeutics, we previously generated RFX001.D, a d-protein antagonist of natural vascular endothelial growth factor A (VEGF-A) that inhibited binding to its receptor. However, RFX001.D is unstable at physiological temperatures (Tm = 33 °C). Here, we describe RFX037.D, a variant of RFX001.D with extreme thermal stability (Tm > 95 °C), high affinity for VEGF-A (Kd = 6 nM), and improved receptor blocking. Comparison of the two enantiomeric forms of RFX037 revealed that the d-protein is more stable in mouse, monkey, and human plasma and has a longer half-life in vivo in mice. Significantly, RFX037.D was nonimmunogenic in mice, whereas the l-enantiomer generated a strong immune response. These results confirm the potential utility of synthetic d-proteins as alternatives to therapeutic antibodies.

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Authors: Uppalapati, M., Lee, D. J., Mandal, K., Li, H., Miranda, L. P., Lowitz, J., Kenney, J., Adams, J. J., Ault-Riché, D., Kent, S. B. H., & Sidhu, S. S.
Affiliations: University of Toronto (CA)
University of Chicago (US)
Amgen (United States) (US)
Antibody Solutions , , ,
Reflexion Pharmaceuticals, Inc. , , ,
Corresponding author: Kent, Stephen B. H.
Sidhu, Sachdev S.
ORCID: Uppalapati, Maruti
Mandal, Kalyaneswar
Li, Hongyan
Lowitz, Joshua
Adams, Jarrett J.
Kent, Stephen B. H.
Sidhu, Sachdev S.
Venue: ACS Chemical Biology
Publisher: American Chemical Society (ACS)
Published: 2016-01-08
Volume: 11
Issue: 4
Pages: 1058-1065
ISSN: 1554-8929 (ISSN-L)
1554-8937
DOI: 10.1021/acschembio.5b01006
OpenAlex: W2238036732
Cited by: 113
References: 45
Citations by year: 2026: 9 · 2025: 17 · 2024: 21 · 2023: 6 · 2022: 7 · 2021: 11 · 2020: 11 · 2019: 9 · 2018: 8 · 2017: 9 · 2016: 5
FWCI: 5.55
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: other-oa
pubs.acs.org/page/policy/authorchoice_termsofuse.html (vor, 26 d)
Locations: ACS Chemical Biology (publishedVersion, other-oa)
PubMed (publishedVersion)
OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
Funders: Amgen (NA)
Basic Energy Sciences (DE-AC02-06CH11357)
Reflexion Pharmaceuticals, Inc. (NA)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Angiogenesis and VEGF in Cancer
Keywords: D-proteins, thermal stability, pharmacokinetics, plasma stability, VEGF receptor binding, enantiomers, synthetic polypeptides
MeSH terms: Calibration, Circular Dichroism, Humans, Reference Standards, Mass Spectrometry, Surface Plasmon Resonance, Vascular Endothelial Growth Factor A

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A Potent d -Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

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